目的:探讨慢病毒介导的CXCR7-shRNA转染人结肠癌细胞株SW620后对CXCR7蛋白表达的影响。方法:(1)设计并合成CXCR7的3对shRNA序列及1对阴性对照序列,与pSilencerTM4.1系统合成构建重组慢病毒载体,转染HEK293T细胞包装病毒并检测滴度;(2)将3种重组慢病毒载体及阴性对照分别感染人结肠癌细胞SW620,RT-PCR检测CXCR7 mRNA的表达情况,测定沉默效率,筛选沉默效率最高的一组CXCR7-shRNA作为后续实验表达载体;(3)MTT法检测转染CXCR7-shRNA对SW620细胞生长增殖的影响;(4)通过细胞划痕实验检测转染CXCR7-shRNA对SW620细胞侵袭迁移能力的影响;(5)Western blot检测转染CXCR7-shRNA对SW620细胞蛋白表达情况。结果:(1)测序证实3对慢病毒载体及1对阴性对照载体均包装成功,滴度分别为3.16×108TU/ml、4.27×108TU/ml、3.93×108TU/ml和2.95×108TU/ml;(2)3组慢病毒载体转染SW620细胞后,CXCR7 mRNA的表达量均较阴性对照组明显降低(P<0.05),其中CXCR7-shRNA-1对CXCR7的抑制率明显高于其他两组(P<0.05);(3)CXCR7-shRNA-1转染SW620后,肿瘤细胞的增殖程度显著减少,与空白组相比有显著性差异(P<0.05);(4)SW620细胞在划痕24h后,空白对照组和实验组的细胞迁移指数(MI)分别为(49.92±6.41)%和(29.13±5.38)%,有统计学意义(P<0.05),划痕48h后,对照组与实验组的MI分别为(96.52±7.44)%和(72.03±8.29)%,有统计学意义(P<0.05);(5)CXCR7-shRNA-1转染SW620细胞后,与空病毒载体组、空白组相比,CXCR7蛋白表达量明显降低,具有统计学意义(P<0.05)。结论:CXCR7-shRNA慢病毒表达载体转染SW620细胞后可有效下调CXCR7 mRNA和蛋白的表达水平并能够抑制肿瘤细胞的增殖与迁移,为下一步研究以CXCR7/CXCL12生物学轴为靶点的结直肠癌慢病毒基因沉默治疗打下了良好的基础,为结直肠癌的基因治疗提供了新方向。 Objective: To investigate the effect of lentivirus-mediated CXCR7-shRNA on CXCR7 expression in human colon cancer cell line SW620. Methods: (1) Three pairs of shRNA sequences of CXCR7 and a pair of negative control sequences were designed and synthesized. Recombinant lentiviral vector was synthesized with pSilencerTM4.1 system and transfected into HEK293T cells to package the virus for titer detection. (2) The recombinant lentiviral vector and negative control were respectively transfected into human colon cancer cell line SW620. The expression of CXCR7 mRNA was detected by RT-PCR and the silencing efficiency was determined. A group of CXCR7-shRNA with the highest silencing efficiency was selected as the subsequent experimental expression vector. (3) The effect of transfection of CXCR7-shRNA on the growth and proliferation of SW620 cells was detected by cell scratch assay; (4) The effect of transfection of CXCR7-shRNA on the invasion and migration of SW620 cells was detected by cell scratch assay; (5) Cell protein expression. Results: (1) Sequencing confirmed that 3 pairs of lentiviral vector and 1 pair of negative control vector were successfully packaged with titer of 3.16 × 108TU / ml, 4.27 × 108TU / ml, 3.93 × 108TU / ml and 2.95 × 108TU / ml, (2) The expression of CXCR7 mRNA in SW620 cells transfected with the three lentiviral vectors was significantly lower than that in the negative control group (P <0.05), and the inhibition rate of CXCR7 by CXCR7-shRNA-1 was significantly higher than that of the other two groups P <0.05). (3) After SW620 was transfected with CXCR7-shRNA-1, the proliferation of tumor cells was significantly reduced compared with the blank group (P <0.05); (4) (49.92 ± 6.41)% and (29.13 ± 5.38)% respectively in the blank control group and the experimental group, with statistical significance (P <0.05). After scratching for 48h, the control group and the experimental group (P <0.05). (5) CXCR7-shRNA-1 transfected SW620 cells, with empty vector group, blank Compared with the control group, CXCR7 protein expression was significantly decreased (P <0.05). CONCLUSION: The CXCR7-shRNA lentiviral vector transfected SW620 cells can effectively down-regulate the expression of CXCR7 mRNA and protein, and can inhibit the proliferation and migration of tumor cells. To further study the effect of CXCR7 / CXCL12 biological axis Rectal cancer lentiviral gene silencing therapy has laid a good foundation for the gene therapy of colorectal cancer provides a new direction.